Snake Venom Gland Organoids

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Snake Venom Gland Organoids
 
Post, Yorick; Puschhof, Jens; Beumer, Joep; Kerkkamp, Harald M.; de Bakker, Merijn A.G.; Slagboom, Julien; de Barbanson, Buys; Wevers, Nienke R.; Spijkers, Xandor M.; Olivier, Thomas; Kazandjian, Taline D.; Ainsworth, Stuart; Iglesias, Carmen Lopez; van de Wetering, Willine J.; Heinz, Maria C.; van Ineveld, Ravian L.; van Kleef, Regina G.D.M.; Begthel, Harry; Korving, Jeroen; Bar-Ephraim, Yotam E.; Getreuer, Walter; Rios, Anne C.; Westerink, Remco H.S.; Snippert, Hugo J.G.; van Oudenaarden, Alexander; Peters, Peter J.; Vonk, Freek J.; Kool, Jeroen; Richardson, Michael K.; Casewell, Nicholas R.; Clevers, Hans
(2020) Cell, volume 180, issue 2, pp. 233 - 247.e21
(Article)
Abstract
Wnt dependency and Lgr5 expression define multiple mammalian epithelial stem cell types. Under defined growth factor conditions, such adult stem cells (ASCs) grow as 3D organoids that recapitulate essential features of the pertinent epithelium. Here, we establish long-term expanding venom gland organoids from several snake species. The newly assembled transcriptome ... read more
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Keywords: heterogeneity, knock-in reporter, Lgr5, organoid, single cell RNA sequencing, snake, stem cells, transcriptomics, venom gland, General Biochemistry,Genetics and Molecular Biology
DOI: https://doi.org/10.1016/j.cell.2019.11.038
ISSN: 0092-8674
Publisher: Cell Press
Note: Funding Information: We thank Reinier van der Linden for flow cytometry assistance; Benedetta Artegiani and Delilah Hendriks for generation of CRISPR-HOT technology and assistance in applying it to snake venom gland organoids; Anko de Graaff and the Hubrecht Imaging Centre (HIC) for microscopy assistance; BaseClear B.V. for bulk mRNA sequencing and de novo transcriptome assembly; Bas Ponsioen for providing the lentiviral H2B-RFP construct; Single Cell Discoveries for the provided single-cell sequencing service and support; Jeremie Tai-A-Pin and Harold van der Ploeg for donating venom gland material to this study; and Sebastiaan Voskuil, Edwin Boel, Marc Bonten, and Frank Driehuis for advice regarding antibiotic treatments. X.M.S. was supported by ALS Foundation Netherlands . N.R.C. was supported by a Sir Henry Dale Fellowship ( 200517/Z/16/Z ) jointly funded by the Wellcome Trust and the Royal Society . Funding Information: We thank Reinier van der Linden for flow cytometry assistance; Benedetta Artegiani and Delilah Hendriks for generation of CRISPR-HOT technology and assistance in applying it to snake venom gland organoids; Anko de Graaff and the Hubrecht Imaging Centre (HIC) for microscopy assistance; BaseClear B.V. for bulk mRNA sequencing and de novo transcriptome assembly; Bas Ponsioen for providing the lentiviral H2B-RFP construct; Single Cell Discoveries for the provided single-cell sequencing service and support; Jeremie Tai-A-Pin and Harold van der Ploeg for donating venom gland material to this study; and Sebastiaan Voskuil, Edwin Boel, Marc Bonten, and Frank Driehuis for advice regarding antibiotic treatments. X.M.S. was supported by ALS Foundation Netherlands. N.R.C. was supported by a Sir Henry Dale Fellowship (200517/Z/16/Z) jointly funded by the Wellcome Trust and the Royal Society. Y.P. J.P. J.B. and H.C. conceptualized the project, designed the experiments, interpreted the results, and wrote the manuscript. H.B. and J. Korving performed immunohistochemistry experiments. J.P. B.d.B. A.v.O. M.C.H. and H.J.G.S. performed bulk and single-cell mRNA sequencing analysis on de novo transcriptome. Y.E.B.-E. assisted with FACS experiments. C.L.I. W.J.v.d.W. and P.J.P. performed transmission electron microscopy. M.A.G.d.B. performed in situ hybridization experiments. R.L.v.I. and A.C.R. generated immunofluorescent images. J.S. and J. Kool performed mass spectrometry analysis. S.A. T.D.K. and N.R.C. performed toxin gene expression analysis on bulk RNA sequencing data. N.R.W. X.M.S. and T.O. performed and analyzed toxicity tests on Mimetas OrganoPlate. H.M.K. F.J.V. and M.K.R. assisted in initializing the project and provided access to venom gland tissue. R.H.S.W. and R.G.D.M.v.K performed and analyzed toxicity tests on rat cortical neurons in MEA plates. W.G. and N.R.C. provided venom gland tissue. H.C. is inventor on several patents related to organoid technology; his full disclosure is given at https://www.uu.nl/staff/JCClevers/. N.R.W. and T.O. are employees of MIMETAS BV, the Netherlands, which is marketing the OrganoPlate. OrganoPlate is a registered trademark of MIMETAS. Publisher Copyright: © 2019 Elsevier Inc.
(Peer reviewed)